Gene therapy-for live cirrhosis by multiplicable hepatocyte and antisense of collagenase-mRNA

【目的】本研究的目的是建立一种增殖性肝细胞移植和给予胶原酶mRNA反义的治疗肝硬化的方法。【材料与方法】研究过程如下：A)建立小鼠和大鼠肝硬化模型，分析肝纤维化的发生机制。B)可增殖的肝细胞杂交瘤由胚胎肝细胞、实质肝细胞和非实质肝细胞组成。C)将杂交瘤移植到肝硬化肝脏中，同时将胶原mRNA的反义蛋白注射到肝脏中。研究了正常肝脏再生的条件。[结果]用CCl_4诱导大鼠肝硬化，并检测肝硬化变化过程中羟丙利酯的活性。并研制了一种新型的激光选择性细胞融合装置。利用该装置制备了具有增殖能力的实质肝细胞和胚胎肝细胞杂交瘤。利用仙台病毒和磷脂制备HVJ脂质体，转染反义胶原酶mrna。在肝纤维化过程分析方面的调查研究仍然存在。

[Purpose] The purpose of this study is to establish a therapy for the liver cirrhosis by a transplantation of proliferative hepatocyte and administration of antisense for mRNA of collagenase. [Materials and Methods] Process of the study were as followed. A) Model of liver cirrhosis is made in mouse and rat and mechanism of fibrosis is analyzed. B) Hybridomas of hepatic cells which can proliferate are made with embrional hepatocytes and parenchymal and nonparenchymal hepatocytes. C) The hybridomas are transplanted into the cirrhotic liver and antisense for the collagenese mRNA is also injected the liver. Conditions of regenerating normal liver are studied. [Results] We made liver cirrhosis using CCl_4 and examined hidroxyprolil activity during changing liver cirrhosis in rats. And we developped a new device for selective cell fusion by Laser device. Using this device we tried to make hybridoma of parenchymal hepatocyte and embrional hepatocyte, which can proliferate. We prepared HVJ liposome using Sendai virus and phospholipid for transfection of antisense of collagenase-mRNA.Studies of investigation in analysis of fibrosing process of liver were still remained.